Purified Kly18 was coated in Nunc Maxisorp plates at 5 μg/ml for one hour at 37 degrees. Wells were emptied and blocked for one hour with 200 μl 1 % skimmed milk in PBS (20 mM sodium phosphate, 150 mM NaCl, pH 7.4). Wells were washed with PBS containing 0.05 % Tween20 (PBS-T) four times. Next, wells were incubated with affinity-purified anti-kly18 IgY diluted in PBS-T for one hour. Wells were washed as above, and anti-IgY antibody was added (Rabbit anti-chicken IgY-HRP, Sigma-Aldrich, A9046, diluted 1/1000 in in PBS-T) for one hour. The ELISA was developed using 3,3’,5,5’-Tetramethylbenzidine (TMB, Sigma-Aldrich) as the substrate with acid stop (read at 450 nm).
Rabbit anti chicken igy hrp
Manufactured by Merck Group
Rabbit anti-chicken IgY-HRP is a laboratory reagent that contains antibodies produced in rabbits that recognize and bind to chicken immunoglobulin Y (IgY) molecules. The antibodies are conjugated with horseradish peroxidase (HRP), an enzyme that can be used for detection in various immunoassay applications.
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2 protocols using rabbit anti chicken igy hrp
1
Kly18 Protein Characterization by ELISA
2
Immunofluorescence Staining of PLVAP
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Mouse anti-human PLVAP mAb9 (link) (clone PAL-E, cat. no. ab8086), monoclonal antibody PAL-E specific for endothelium, and mouse anti-human Plvap mAb Clone 174/210 (link) were purchased from Abcam (Cambridge, MA). Hybridoma cells secreting PAL-E mAb was obtained from M. Kahn (University of Pennsylvania) and was used to produce PAL-E mAb by BioXCell (Lebanon, NH). Anti-HA mAb clone HA.11 was used (Covance, Princeton, NJ). Chicken anti-human PV1C pAb was previously described elsewhere.8 (link) Goat anti-mouse IgG-HRP was obtained from Biodesign (Saco, ME), rabbit anti-chicken IgY-HRP was obtained from Sigma-Aldrich (St. Louis, MO), and goat anti-mouse IgG-AlexaFluor 647 was obtained from Life Technologies (Eugene, OR).
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